1. Field of the Invention
This invention relates to an improvement in promoting the rate of association for high specificity binding pairs used in a variety of industrial, research and medical applications. These pairs include enzyme/substrate, complementary polynucleotide and antibody/antigen combinations. In one specific embodiment, this invention relates to the acceleration of nucleic acid hybridization by heterogeneous nuclear ribonucleoproteins [hnRNPs]. In another specific embodiment, this invention relates to the acceleration of nucleic acid hybridization by a cationic detergent.
2. Information Disclosure
The acceleration of annealing between complementary nucleic acids has been described. Christiansen C. and Baldwin, R. L., 1977, Catalysis of DNA Reassociation by the Escherichia coli DNA Binding Protein, J. Mol. Biol. 115:441-454; Weinstock, G. M. et al., 1979, ATP-dependent renaturation of DNA catalyzed by the recA protein of Escherichia coli, Proc. Natl. Acad. Sci. 76:126-130; Cox, M. M. and Lehman, I. R., 1981, Renaturation of DNA: a novel reaction of histones, Nucleic Acid research 9:389-399; Keener S. L. and McEntree, K., 1984, Homologous pairing of single-stranded circular DNAs catalyzed by recA protein, Nucleic Acids Research 12:6127-6139; and Bryant, F. R. et al., 1989, Kinetic Modeling of the RecA Protein Promoted Renaturation of Complementary DNA Strands, Biochemistry 28:1062-1069.
Heterogeneous nuclear particles was known and reviewed by Dreyfuss, G., et al., March 1988, Heterogeneous nuclear ribonucleoprotein particles and the pathway of mRNA formation, TIBS 13:86-90 and Bandziulis, R. J. et al., 1989, RNA-binding proteins as developmental regulators, Genes & Devel. 3:431-437.
The A1 core protein has been implicated in helix-destabilization. Williams, K. R. et al., 1985, Amino acid sequence of the UP1 calf thymus helix-destabilizing protein and its homology to an analogous protein from mouse myeloma, Proc. Natl. Acad. Sci. USA 82:5666-5670. The cDNA encoding A1 hnRNP from rat has been cloned and expressed. Cobianchi, F. et al., 1986, Structure of Rodent Helix-destabilizing Protein Revealed by cDNA Cloning, J of Biol. Chem. 261:3536-3543. The A1 hnRNP from human cells has been isolated and purified. Kumar, A. et al., 1986, Purification and Domain Structure of Core hnRNP Proteins A1 and A2 and Their Relationship to Single-stranded DNA-Binding Proteins, J. Biol. Chem. 261:11266-11273. Kumar et al also reported on the ability of A1 hnRNP to mediate duplex formation between synthetic polynucleotides.
The characterization of mammalian A1 hnRNP was described by Cobianchi, F. et al. 1988, Mammalian Heterogeneous Nuclear Ribonucleoprotein Complex Protein A1, J. Biol. Chem. 263:1063-1071 and by Merrill B. M. et al., 1988, Phenylalanines That Are Conserved among Several RNA-binding Proteins Form Part of A Nucleic Acid-binding Pocket in the A1 Heterogeneous Nuclear Ribonucleoprotein, J. Biol. Chem. 263:3307-3313.